Quantitative proteomics analysis of tomato growth inhibition by ammonium nitrogen

Abstract

As a single nitrogen source, ammonium (NH₄⁺) can inhibit the growth of plants, especially when applied in excess. Tandem mass tag (TMT) quantitative proteomics technology was employed in the current study to explore and analyze the mechanisms of ammonium-induced inhibition. F₁ tomato (Lycopersicon esculentum Mill) was used in this study. Seedlings at the four leaf-stages grown in a greenhouse were irrigated using nutrient solution with NH₄⁺-N as single nitrogen source (15 mmol L^-1, single NO₃^--N as control) for 5 weeks. Compared to the control, the root biomass of NH₄⁺-N-treated seedlings decreased by 50%. In addition, NH₄⁺ content in roots was 2.83-fold increased and soluble sugar and protein contents were increased. However, the starch content did not change significantly. The activities of glutamine synthetase (GS), glutamate synthetase (GOGAT) and glutamate dehydrogenase (GDH), which are involved in ammonium assimilation, were increased, and glutamine (Gln) content was also increased. However, glutamate (Glu) content, which is important for amino transfer, did not significantly increase. Ammonium assimilation was inhibited. Root quantitative proteomics showed that carbonic anhydrase Q5NE21 was significantly downregulated. Although K4BPV5 and K4D9J3 proteins, which improve ammonium assimilation, were upregulated, ammonium assimilation was limited. In addition, NH₄⁺ accumulated, which is likely due to Q5NE21 downregulation. Meanwhile, cell wall metabolism related to phenylpropanoid biosynthesis was altered due to the accumulation of NH₄⁺ levels. Subsequently, tomato root growth was inhibited.

Keywords: Ammonium nitrogen; Growth inhibition; Proteomics; Tomato.